Molecular cloning of human intestinal mucin cDNAs. Sequence analysis and evidence for genetic polymorphism

• Published in: The Journal of biological chemistry Vol. 264; no. 11; pp. 6480 - 6487
• Main Authors: GUM, J. R, BYRD, J. C, HICKS, J. W, TORIBARA, N. W, LAMPORT, D. T. A, KIM, Y. S
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 15.04.1989
• Subjects: Amino Acid Sequence; Base Sequence; Biological and medical sciences; Blotting, Northern; Blotting, Southern; Blotting, Western; Cloning, Molecular; DNA - genetics; Fundamental and applied biological sciences. Psychology; genes; Genes. Genome; Humans; intestine; Intestines; man; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; mucin; Mucins - genetics; Mucins - immunology; Polymorphism, Genetic; Protein Biosynthesis; Human; Molecular hybridization; Nucleotide sequence; Antibody; Gut; Gene library; Tandemly repeated sequence; Screening; Mucin; Complementary DNA; Immunoblotting assay; Aminoacid sequence; Polymorphism
• ISSN: 0021-9258; 1083-351X

A human small intestine lambda gt11 cDNA library was screened using antisera prepared against the deglycosylated protein backbone of human colon cancer xenograft mucin. Three cDNAs were isolated from this screening, designated SMUC 40-42. These cDNAs were all found to contain tandem repeats of 69 nucleotides which encoded a threonine- and proline-rich protein consensus sequence of PTTTPITTTTTVTPTPTPTGTQT. RNA blots probed with one of these cDNAs, SMUC 41, exhibited large, polydisperse hybridization bands at approximately 7,600 bases. Band intensities were strongest when human small intestine, colon, and colon cancer poly(A)+ RNA was used. In vitro translation of poly(A)+ RNA from human small intestine, colon, and colon cancer cells produced a 162,000-dalton peptide that was immunoprecipitated with antibodies to deglycosylated mucin. SMUC 41 was also used to probe DNA blots, which indicated the presence of restriction fragment length polymorphisms in the intestinal mucin gene. These findings may be important in assessing the abnormal mucins found associated with several human diseases.